Targeted chondroitinase abc fusion proteins and complexes thereof

Abstract

Described herein are targeted ChABC fusion proteins, complexes thereof, and uses thereof. The targeted ChABC fusion proteins can include a ChABC polypeptide that can be linked to a Gal-3 polypeptide. Monomer targeted ChABC fusion proteins can form homogeneous or heterogeneous complexes. The targeted ChABC fusion proteins and complexes thereof can be formulated as pharmaceutical formulations. The targeted ChABC fusion proteins, complexes thereof, and formulations thereof can be administered to a subject in need thereof.

We claim: 
     
         1 . A targeted effector fusion protein comprising:
 a chondroitinase ABC (ChABC) polypeptide; and   a galectin-3 (Gal-3) polypeptide, wherein the Gal-3 is operatively linked to the ChABC polypeptide.   
     
     
         2 . The targeted effector fusion protein of  claim 1 , wherein the ChABC polypeptide is directly fused to the Gal-3 polypeptide. 
     
     
         3 . The targeted effector fusion protein of  claim 1 , wherein the ChABC polypeptide is operatively linked to the Gal-3 polypeptide via a linker. 
     
     
         4 . The targeted effector fusion protein of  claim 3 , wherein the linker is a flexible linker or a rigid linker. 
     
     
         5 . The targeted effector fusion protein of  claim 4 , wherein the flexible linker is selected from the group consisting of: a peptide, a polypeptide, a cross-linking reagent, and a coupling agent. 
     
     
         6 . The targeted effector fusion protein of any one of  claims 1 - 5 , wherein the Gal-3 is capable of specifically binding to a carbohydrate. 
     
     
         7 . The targeted effector fusion protein of any one of  claims 3 - 6 , wherein the linker is an alpha coil polypeptide or a random coil polypeptide. 
     
     
         8 . The targeted effector fusion protein of  claim 7 , wherein the alpha coil polypeptide comprises one or more heptads, where each heptad has the general formula of A-B-C-D-E-F-G, wherein amino acids A and D are each a hydrophobic amino acid, wherein amino acids B, C, E, F, and G, are each independently selected from a hydrophilic amino acid, a polar amino acid, or a charged amino acid. 
     
     
         9 . The targeted effector fusion protein of any one of  claims 3 - 8 , wherein the linker comprises a polypeptide sequence that is about 90 to 100% identical to any one of SEQ ID NOs: SEQ ID NO: 17-32. 
     
     
         10 . The targeted effector fusion protein of any one of  claims 7 - 8 , wherein the alpha coil polypeptide is capable of multimerizing with one or more other alpha coils that are integrated in one or more other targeted effector fusion proteins. 
     
     
         11 . The targeted effector fusion protein of  claim 1 , wherein the targeted effector fusion protein comprises a polypeptide sequence that is about 90% to 100% identical to any one of SEQ ID NOs: 15-16. 
     
     
         12 . A multimeric targeted effector fusion protein complex comprising:
 at least two targeted effector fusion proteins, wherein each of the at least two targeted effector proteins is according to any one of  claims 7 - 11 , and the at least two targeted effector proteins are conjugated to each other by binding between the alpha coil polypeptide or random coil polypeptide in each of the at least two targeted effector proteins.   
     
     
         13 . The multimeric targeted effector fusion protein complex of  claim 12 , wherein the multimeric targeted effector fusion protein complex is homogeneous. 
     
     
         14 . The multimeric targeted effector fusion protein complex of  claim 12 , wherein the multimeric targeted effector fusion protein complex is heterogeneous. 
     
     
         15 . A single fusion polypeptide sequence comprising:
 a chondroitinase ABC (ChABC) polypeptide; and   a galectin-3 (Gal-3) polypeptide, wherein the Gal3 is operatively linked to the effector protein via a linker.   
     
     
         16 . A single fusion polypeptide sequence comprising:
 a chondroitinase ABC (ChABC) polypeptide; and   a galectin-3 (Gal-3) polypeptide, wherein the Gal3 is directly fused to the ChABC polypeptide.   
     
     
         17 . A polynucleotide that encodes any one of the targeted effector fusion proteins in any one of  claims 1 - 11  or  15 - 16 . 
     
     
         18 . The polynucleotide of  claim 17 , wherein the polynucleotide has a sequence that is abbot 50% to 100% identical to any one of SEQ ID NOs: 6-7 
     
     
         19 . A vector comprising:
 a polynucleotide according to any one of  claims 17 - 18 .   
     
     
         20 . A single fusion polypeptide sequence comprising:
 at least two targeted effector fusion proteins as in any one of  claims 7 - 11 , wherein each of the at least two targeted effector fusion proteins are directly fused at the C-terminus, N-terminus, or both the C-terminus and N-terminus of at least one other targeted effector fusion protein of the at least two targeted effector fusion proteins.   
     
     
         21 . A single fusion polypeptide sequence comprising:
 at least two targeted effector fusion proteins as in any one of  claims 7 - 11 , wherein each of the at least two targeted effector fusion proteins are operatively linked at the C-terminus, N-terminus, or both the C-terminus and N-terminus to at least one other targeted effector fusion protein of the at least two targeted effector fusion proteins via one or more additional amino acids.   
     
     
         22 . A polynucleotide encoding the single fusion polypeptide as in any one of  claim 20  or  21 . 
     
     
         23 . A vector comprising:
 the polynucleotide according to  claim 22 .   
     
     
         24 . A pharmaceutical formulation comprising:
 a targeted effector fusion protein as in any one of  claims 1 - 11  or  15 - 16 ; and   a pharmaceutically acceptable carrier.   
     
     
         25 . A pharmaceutical formulation comprising:
 a multimeric targeted effector fusion protein complex as in any one of  claims 12 - 14 ; and   a pharmaceutically acceptable carrier.   
     
     
         26 . A method comprising:
 administering a targeted effector fusion protein as in any one of  claims 1 - 11  or  15 - 16  or a targeted effector fusion protein complex as in any one of  claims 12 - 14  to a subject.   
     
     
         27 . The method of  claim 29 , wherein the subject has a disease. 
     
     
         28 . The method of any one of  claims 26 - 27 , wherein the subject has a central nervous system injury or symptom thereof. 
     
     
         29 . The method of any one of  claims 26 - 27 , wherein the subject has glial scarring. 
     
     
         30 . The method of any one of  claims 26 - 27 , wherein the subject has a ChABC deficiency. 
     
     
         31 . The method of any one of  claims 26 - 27 , wherein the subject is in need of neuron regeneration. 
     
     
         32 . The method of any one of  claims 26 - 31 , wherein the administration is local.